Multiplex Assay Kit for Heparin Binding Epidermal Growth Factor Like Growth Factor (HBEGF) ,etc. by FLIA (Flow Luminescence Immunoassay)

HB-EGF; DTR; DTS; DTSF; HEGFL; Diphtheria Toxin Receptor; Proheparin-binding EGF-like growth factor; Diphtheria toxin receptor

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Heparin Binding Epidermal Growth Factor Like Growth Factor (HBEGF) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Heparin Binding Epidermal Growth Factor Like Growth Factor (HBEGF) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Heparin Binding Epidermal Growth Factor Like Growth Factor (HBEGF) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Heparin Binding Epidermal Growth Factor Like Growth Factor (HBEGF) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 99-105 102
EDTA plasma(n=5) 87-101 95
heparin plasma(n=5) 79-90 82
sodium citrate plasma(n=5) 80-101 81

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Heparin Binding Epidermal Growth Factor Like Growth Factor (HBEGF) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Heparin Binding Epidermal Growth Factor Like Growth Factor (HBEGF) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Heparin Binding Epidermal Growth Factor Like Growth Factor (HBEGF) ,etc. by FLIA (Flow Luminescence Immunoassay) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 98-105% 81-88% 95-103% 86-101%
EDTA plasma(n=5) 93-105% 93-105% 90-98% 80-89%
heparin plasma(n=5) 92-105% 85-98% 86-95% 95-103%
sodium citrate plasma(n=5) 96-105% 96-104% 80-88% 88-97%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:HBEGF) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
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European Journal of Pain A novel method for assessing bladder-related pain reveals the involvement of nerve growth factor in painassociated with cyclophosphamide-induced chronic cystitis in mice. Pubmed:25820250
Kinki daigaku 2 型糖尿病モデルラットにおける糖尿病性腎症の進行とHB-EGF の関連性 17844
laboratory investigation Heparin-binding epidermal growth factor contributes to COPD disease severity by modulating airway fibrosis and pulmonary epithelial–mesenchymal transition Pubmed:29581578
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JOURNAL OF CELLULAR AND MOLECULAR MEDICINE Polarization of ADAM17‐driven EGFR signalling in electric field‐guided collective migration of epidermal sheets Pubmed: 33164313
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